信阳水牛肌纤维类型调控相关基因组织表达及分化时序研究

doi:10.19543/j.cnki.1673-4556.20260416.001

Abstract

Abstract:

Objective To elucidate the functions of MSTN, BDNF, and DKK3 genes associated with slow-to-fast muscle fiber conversion in Xinyang buffalo and their effects on meat quality characteristics, in order to provide reference for the improvement of Xinyang buffalo breeds. Method Three muscle fiber-related genes (MSTN, BDNF, DKK3) were selected from relevant literature. Their amino acid sequences were downloaded from NCBI, domain prediction was performed using MEME, and protein interaction analysis was conducted through the online software STRING for bioinformatics evaluation. Tissue samples from three 36-month-old bulls (Liver, spleen, lung, kidney, heart, foreleg muscle, hindleg muscle, longissimus dorsi muscle, subcutaneous muscle, large intestine, small intestine) were collected for tissue expression profiling. Collect the longissimus dorsi muscle from a 1-day-old newborn calf, isolate and culture the primary cells, induce their differentiation, and analyze time-course gene expression. Result Bioinformatics analysis reveals that the protein encoded by the MSTN gene contains domains associated with the TGF-β signaling pathway and is predominantly highly expressed in skeletal muscle tissue. The protein encoded by the BDNF gene possesses domains related to neurotrophic factors and is primarily highly expressed in neural tissues and skeletal muscle. The protein encoded by the DKK3 gene contains domains associated with the Wnt signaling pathway and is mainly highly expressed in adipose and liver tissues. GO and KEGG analyses further indicated that these genes were respectively enriched in the TGF-β signaling pathway, neurotrophic factor signaling pathway, and Wnt signaling pathway. The temporal differentiation results showed that the expression of MSTN was upregulated in the early stage of differentiation and then rapidly downregulated; the expression level of the BDNF gene exhibited a gradual upward trend and was reactivated and upregulated at the late differentiation stage; DKK3 was not expressed during the differentiation of primary myoblasts. Conclusion These results indicate that the expression of MSTN, BDNF, and DKK3 genes is associated with the formation of muscle fibers in buffalo.

Key words: Xinyang buffalo, MSTN, BDNF, DKK3, Bioinformatics analysis, Tissue expression

CLC Number:

S823

Xiaoge Zhang, Yanduo Zhou, Jianzhang Li, Luping Ma, Jun Li, Ruijie Hao, Zijing Zhang, Yun Ma, Tianliu Zhang, Chengcheng Liang. Tissue expression and differentiation characteristics of genes regulating muscle fiber types in Xinyang buffalo[J].Chinese Livestock and Poultry Breeding, 2026, 22(5): 32-42.

Figures/Tables 8

Table 1

Fig. 1

Fig. 2

Fig. 3

Table 2

Table 3

Protein interaction network GO enrichment analysis"

项目 Items	基因 Genes	项目ID Term ID	项目描述 Term description	基因数量 Gene count	错误率FDR	基因 Genes
细胞组分 Cellular component	MSTN	GO: 0048179	激活素受体复合体	4	1.99E-08	TGFBR1, ACVR2A, ACVR1B, ACVR2B
	BDNF	GO: 0045202	突触	5	0.0174	BDNF, NGFR, NTRK2, NTF4, NGF
	BDNF	GO: 0120025	质膜界定的细胞突起	6	0.0174	CNTF, BDNF, NTRK2, NTF4, NGF, GFAP
	DKK3	GO: 0005576	细胞外区域	8	0.0014	DKK3, FRZB, DKK2, WIF1, SFRP4, SFRP2, SFRP1, DKK1
分子功能 Molecular function	MSTN	GO: 0048179	激活素受体复合体	4	1.99E-08	TGFBR1, ACVR2A, ACVR1B, ACVR2B
	MSTN	GO: 0048185	激活素结合	6	2.20E-13	FST, TGFBR1, ACVR2A, ACVR1B, FSTL3, ACVR2B
	BDNF	GO: 0017002	激活素受体活性	4	4.83E-08	TGFBR1, ACVR2A, ACVR1B, ACVR2B
	DKK3	GO: 0017147	Wnt蛋白结合	5	1.02E-08	LRP5, FRZB, SFRP4, SFRP2, SFRP1
		GO: 0005102	信号受体结合	6	0.0105	DKK3, DKK2, WIF1, SFRP2, SFRP1, DKK1
		GO: 0005515	蛋白质结合	10	0.021	DKK3, LRP5, FRZB, DKK2, WIF1, SFRP4, CTNNB1, SFRP2, SFRP1, DKK1
		GO: 0030545	信号受体调节活性	4	0.0462	DKK3, DKK2, SFRP2, DKK1
		GO:0017147	Wnt蛋白结合	5	1.02E-08	LRP5, FRZB, SFRP4, SFRP2, SFRP1
生物学过程Biological process	MSTN	GO: 0032925	激活素受体信号通路的调控	5	6.04E-09	FST, SMAD2, ACVR2A, ACVR1B, FSTL3
		GO: 0007178	跨膜受体蛋白丝氨酸/苏氨酸激酶信号通路	7	7.99E-09	FST, MSTN, SMAD2, TGFBR1, ACVR2A, ACVR1B, ACVR2B
		GO: 0045595	细胞分化调控	10	7.99E-09	MYOD1, FST, MYOG, MSTN, SMAD2, TGFBR1, ACVR2A, MYF5, ACVR1B, FSTL3
	BDNF	GO: 0038179	神经营养因子信号通路	5	4.78E-09	BDNF, NTRK2, NTF4, NGF, SORT1
	DKK3	GO: 0090090	典型Wnt信号通路的负调控	9	4.90E-15	DKK3, FRZB, DKK2, SFRP4, CTNNB1, KREMEN1, SFRP2, SFRP1, DKK1
	DKK3	GO: 0045597	细胞分化的正调控	7	4.31E-06	LRP5, FRZB, WIF1, SFRP4, CTNNB1, SFRP2, SFRP1

Table 3

Fig. 4

Fig. 5

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基因Genes	引物序列（5'-3'）Primer sequence	产物长度Product length/bp	退火温度Annealing temperature/℃
MSTN-F	CATTACCATGCCCACGGAGTCTG	149	63.6
MSTN-R	TCGCAGGAGTCTTGACAGGTCTC	149	63.6
BDNF-F	AACTCCCAGTGCCGAACTACCC	80	64.1
BDNF-R	ATGAACCGCCAGCCAATACGC	80	64.1
DKK3-F	TCGTCCTCTCGGCAGTGTTACTC	148	63.6
DKK3-R	ACTCGGGCAAGCAAGATGACATC	148	63.6
β- actin-F	CATCCTGACCCTCAAGTA	91	52.3
β -actin-R	CTCGTTGTAGAAGGTGTG	91	52.3

核心蛋白 Core protein	通路ID Term ID	通路描述 Term description	基因数量 Gene count	错误率 FDR	基因 Genes
MSTN	bta04350	TGF-β信号通路	6	7.56E-10	FST, SMAD2, TGFBR1, ACVR2A, ACVR1B, ACVR2B
	bta04550	调节干细胞多能性	5	6.67E-07	SMAD2, ACVR2A, MYF5, ACVR1B, ACVR2B
	bta04060	细胞因子-受体相互作用	5	1.62E-05	MSTN, TGFBR1, ACVR2A, ACVR1B, ACVR2B
BDNF	bta04722	神经营养因子	6	2.24E-09	BDNF, NGFR, NTRK2, NTF4, NGF, SORT1
	bta04151	PI3K-Akt信号通路	6	9.36E-07	BDNF, IL6, NGFR, NTRK2, NTF4, NGF
	bta04014	Ras信号通路	5	5.85E-06	BDNF, NGFR, NTRK2, NTF4, NGF
	bta04010	MAPK信号通路	5	1.03E-05	BDNF, NGFR, NTRK2, NTF4, NGF
DKK3	bta04310	Wnt信号通路	7	6.33E-11	LRP5, DKK2, SFRP4, CTNNB1, SFRP2, SFRP1, DKK1

Tissue expression and differentiation characteristics of genes regulating muscle fiber types in Xinyang buffalo

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